title: Down-regulation of MeCP2 in Hirschsprung's disease.
abstract: OBJECTIVE
  Hirschsprung's disease (HSCR) is a congenital disorder characterized by the absence of intramural ganglion cells which are highly associated with impaired proliferation and migration of neural crest cells. Whether methyl CpG binding protein 2 (MeCP2) is related with HSCR still remains unknown. This study investigates the involvement of MeCP2 in HSCR.
  
  
  METHODS
  Quantitative real time PCR and western blot were used to detect the expression level of MeCP2 both in the aganglionic/diseased segment and the ganglionic/normal segment. In vitro assays we used siRNAs to knock-down the expression of MeCP2 in SH-SY5Y cell lines, and furthermore, MTT and transwell assays were used to detect the proliferation and migration ability, respectively. In addition, bisulfite sequencing (BSP) and miRNA analysis were used to examine why MeCP2 is decreased in HSCR samples.
  
  
  RESULTS
  MeCP2 exhibited a lower expression level in tissues of HSCR patients compared with the controls. The down-regulation may also suppress the proliferative ability of the cells. However, there was no significant difference in the MeCP2 methylation level between cases and controls. Similarly, there was no difference between cases and controls in miRNA-34b (miR-34b) which is predicted to regulate MeCP2 through complementary binding to the 3'-untranslated region of MeCP2.
  
  
  CONCLUSIONS
  Our results indicated that an aberrant decreased level of MeCP2 may play an important role in the pathogenesis of HSCR.
authors.affiliation: State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, China; Department of General Surgery, Yixing Traditional Chinese Medicine Hospital, Wuxi 214200, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Zhou
authors.forename: Jingjing
authors.initials: J
authors.lastname: Qin
authors.forename: Junwei
authors.initials: J
authors.lastname: Tang
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.forename: Qiming
authors.initials: Q
authors.lastname: Geng
authors.forename: Weiwei
authors.initials: W
authors.lastname: Jiang
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Virender
authors.initials: V
authors.lastname: Rehan
authors.forename: Weibing
authors.initials: W
authors.lastname: Tang
authors.forename: Xiaoqun
authors.initials: X
authors.lastname: Xu
authors.forename: Yankai
authors.initials: Y
authors.lastname: Xia
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D016022:Case-Control Studies; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D003106:Colon / Q000378:metabolism*; D019175:DNA Methylation; D015536:Down-Regulation; D005260:Female; D006627:Hirschsprung Disease / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007223:Infant; D008297:Male; D051783:Methyl-CpG-Binding Protein 2 / Q000235:genetics / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012007:Rectum / Q000378:metabolism*
chemical_list: D015415:Biomarkers; C497392:MECP2 protein, human; C522425:MIRN34 microRNA, human; D051783:Methyl-CpG-Binding Protein 2; D035683:MicroRNAs
keywords: BSP; Hirschsprung's disease; MeCP2; Proliferation; miRNA